地衣芽孢杆菌发酵条件优化和抑菌活性测定  

Optimization of Fermentation Conditions and Determination of Antibacterial Activity of Bacillus licheniformis

作　　者：张钰 马曦[2] 赵金标[2] 鲁琳[1] ZHANG Yu;MA Xi;ZHAO Jinbiao;LU Lin(College of Animal Science and Technology,Beijing University of Agriculture,Beijing 102206,China;College of Animal Science and Technology,China Agricultural University,Beijing 100193,China)

机构地区：[1]北京农学院动物科学技术学院,北京102206 [2]中国农业大学动物科学技术学院,北京100193

出　　处：《中国畜牧兽医》2022年第7期2812-2819,共8页China Animal Husbandry & Veterinary Medicine

基　　金：博士后创新人才支持计划(BX20200365);中国农业大学横向项目“高效IGF-1和生物酵母技术开发”(201905410411435)。

摘　　要：【目的】探究不同培养及发酵条件对地衣芽孢杆菌19148抑菌作用的影响及地衣芽孢杆菌19148的耐药性。【方法】设计单因素试验,分别在不同温度、酸碱度、金属离子、摇床转速、发酵时间和接种量的条件下培养地衣芽孢杆菌19148菌液,将培养后的菌液离心、过滤制成无菌滤液,并通过牛津杯抑菌试验探究无菌滤液对大肠杆菌K88、鼠伤寒沙门氏菌和金黄色葡萄球菌1882的抑制能力。通过药敏纸片法探究地衣芽孢杆菌19148对27种抗菌药的耐药性。【结果】地衣芽孢杆菌19148在4和75℃下生长受到抑制,25、37和45℃能正常生长,其中在37℃抑菌活性最好。在pH 5.0~9.0的条件下能正常生长,在pH为6.0条件下抑菌活性最好,对铜离子不耐受,但对镁离子表现出了良好的耐受性,且不同浓度镁离子处理组间抑菌活性无显著差异(P>0.05)。体外发酵试验中,地衣芽孢杆菌19148在3%种子液接种量、150 r/min发酵28 h时,体外发酵的抑菌活性最好。耐药性评价试验结果显示,地衣芽孢杆菌19148对青霉素、四环素、头孢他啶、呋喃唑酮和多黏菌素B 5种抗菌药中介,对头孢哌酮和红霉素等其他22种抗菌药敏感。【结论】地衣芽孢杆菌19148具有较好的稳定性,能应用于工业发酵饲料生产,对抗菌药不具有耐药性。【Objective】The objective of this study was to investigate the effects of different culture and fermentation conditions on the antibacterial ability of Bacillus licheniformis 19148 and the resistance of Bacillus licheniformis 19148.【Method】Bacillus licheniformis 19148 was cultured under different conditions of temperature,pH,metal ions,shaking speed,fermentation time and inoculation amount in a single factor experimental design,respectively,then centrifuge and filter the cultured liquid into sterile filtrate to study inhibition ability against Escherichia coli K88,Salmonella Typhimurium and Staphylococcus aureus 1882 using oxford cup bacteriostatic test.The drug resistance of Bacillus licheniformis 19148 to 27 antibiotics was studied by drug sensitive paper method.【Result】Bacillus licheniformis 19148 could grow normally at 25,37 and 45℃,and was inhibited at 4 and 75℃,and had the best antibacterial activity at 37℃.Bacillus licheniformis 19148 could grow normally in the pH ranged from 5.0 to 9.0 and had the best antibacterial activity at pH 6.0.Bacillus licheniformis 19148 was intolerant to copper ions,but could tolerate magnesium ions,and there was no significant difference among treatment groups with different magnesium ion concentrations.In in vitro fermentation,Bacillus licheniformis 19148 showed the best antibacterial activity under the conditions of 3%seed liquid inoculation,and 150 r/min fermetation for 28 h.In the drug resistance evaluation test,Bacillus licheniformis 19148 showed moderate sensitivity to penicillin,tetracyclines,ceftazidine,furazolidone and polymyxin B,and showed high sensitivity to the other 22 antibiotics such as cefoperazone and erythromycin.【Conclusion】Bacillus licheniformis 19148 had great stability and could be used in the production of industrial fermented feed without antibiotic resistance.

关 键 词：地衣芽孢杆菌 发酵条件 抑菌活性 耐药性 益生菌制剂 

分 类 号：S852.615[农业科学—基础兽医学]